| Assay Method Information | |
| | hERG Qpatch Assay Protocol |
| Description: | hERG currents were recorded using the Qpatch automated patch clamp systems (Sophion Bioscience Inc., North Brunswick, NJ) in the whole (single) cell configuration. hERG expressing CHO-K1 cells were harvested with Detachin (Genlantis) and stored in the modified serum-free SFM-2 media (Life Technologies) at room temperature. The extracellular solution contained (in mM) NaCl (145), KCl (4), MgCl2 (1), CaCl2 (2), and HEPES (10), pH 7.4, with NaOH. The intracellular solution contained KCl (135), MgCl2 (1.75), CaCl2 (5.4), EGTA (10), K2-ATP (4), and HEPES (10), pH 7.2, with KOH. After whole cell configuration was achieved, the cell was held at −90 mV, and a 0.1 s pulse to −50 mV was delivered to measure the leaking current, which was subtracted from the tail current online. Then the cell was depolarized to +20 mV for 4 s (prepulse), followed by a 4 s test pulse to −50 mV to reveal the hERG tail current. To monitor changes in the current amplitude, this voltage protocol was repeatedly applied every 20 s. Test compounds were first diluted in DMSO for six dose-response experiments and then dissolved in the extracellular solution using Freedom EVO liquid handling robotic system (Tecan, M nnedorf, Switzerland). |
| Affinity data for this assay | |
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